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研究生:陳音曲
研究生(外文):Yin-Chiu Chen
論文名稱:MERRF腦肌病症候群患者的皮膚纖維母細胞之抗氧化酵素與間質金屬蛋白酵素-1表現的研究
論文名稱(外文):Studies on Changes in the Expression of Antioxidant Enzymes and Matrix Metalloproteinase-1 in Skin Fibroblasts of Patients with MERRF Syndrome
指導教授:魏耀揮魏耀揮引用關係
指導教授(外文):Yau-Huei Wei
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生物化學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:89
中文關鍵詞:粒線體抗氧化酵素間質金屬蛋白酵素-1
外文關鍵詞:mitochondriaantioxidant enzymesmatrix metalloproteinase-1
相關次數:
  • 被引用被引用:1
  • 點閱點閱:204
  • 評分評分:
  • 下載下載:22
  • 收藏至我的研究室書目清單書目收藏:0
有許多神經肌肉系統相關的疾病常已被證實與粒線體DNA的突變有關。MERRF症候群是其中一個很顯著的例子,大多數的患者病變組織細胞內都攜帶有粒線體DNA上 tRNALys基因的A8344G突變,多年來的研究大都關注於這樣的粒線體DNA突變在此一疾病發生所扮演的角色,但是MERRF的致病機轉至今仍不清楚。過去本實驗室曾利用Agilent human 1 cDNA microarry和RT-PCR分析一個MERRF患者纖維母細胞的基因表現,顯示在MERRF患者的細胞中,其與發炎反應相關之蛋白、重建細胞間質相關酵素(如:MMPs)、及作為氧化壓力指標之蛋白皆有基因活化的現象。在本研究裡,我把焦點放在抗氧化酵素及間質金屬蛋白酵素(MMP)表現與MERRF致病機轉的相關性。分析此家族成員纖維母細胞抗氧化酵素表現,我發現MERRF女性患者之皮膚纖維母細胞的Mn-SOD不論在蛋白質含量抑或酵素活性皆比他人高出許多,而細胞內H2O2的累積也較高。此外,其MMP-1的表現更是顯著的高出數倍以上。而MnSOD、H2O2及MMP-1較高表現之現象也可以在來自其他三株MERRF患者之皮膚纖維母細胞中觀察到。然而,來自另兩位MERRF患者之不帶有A8344G突變纖維母細胞,則測量出其細胞內有大量的超氧陰離子累積。另一方面,實驗結果顯示出細胞分泌的MMP1酵素活性會因外加200□□M H2O2 而有被誘導的現象,反之,若加入1 mM NAC則MMP1的活性顯著地被抑制。本研究結果顯示在MERRF患者之皮膚纖維母細胞內有較高的氧化壓力,以致造成氧化性傷害。此外,我證實患者細胞內MMP-1的表現與H2O2含量息息相關。這些基因表現的改變顯示出粒線體結構分佈的破壞及組織細胞所遭受的氧化傷害,皆在MERRF症候群的發生過程及致病機轉的研究上扮演著極為重要的角色。
Mitochondrial DNA mutations (mtDNA) have been demonstrated to associate with a wide spectrum of neuromuscular disease in humans. A well-studied example is the A8344G mutation in mtDNA that causes myoclonic epilepsy with ragged-red fibers (MERRF) syndrome, whose mechanism of pathogenesis is still not clear. In one of our previous studies, the induction of inflammation-related proteins, matrix metalloproteinases, and markers of oxidative stress have been observed in the patient’s fibroblasts by Agilent human 1 cDNA microarray and RT-PCR. It is of great interest to elucidate the relationship between gene expressions of antioxidant enzymes and MMPs and MERRF syndrome. We analyzed the expression of antioxidant enzymes and found that protein and activity levels of MnSOD were significantly increased in skin fibroblasts of a female patient with MERRF syndrome. The intracellular level of hydrogen peroxide was increased in skin fibroblasts of the MERRF patient as compared to those of her family members and age-matched controls. Moreover, the expression of MMP-1 in skin fibroblasts of the MERRF patient was also higher than that of controls. Increased expression in MnSOD, MMP1 and higher level of hydrogen peroxide were also observed in skin fibroblasts from other three MERRF patients harboring A8344G mutated mtDNA. However, we found that skin fibroblasts from two MERRF patients without the A8344G mutation had higher level of superoxide anion. On the other hand, we found that the expression of MMP1 could be inhibited by treatment with 1 mM N-acetylcysteine and was induced by treatment with 200 □M hydrogen peroxide. The morphology and sub-cellular orgahization of mitochondria in the disrupted network were significantly altered and abnormal dot-like discrete structures disappeared. Taken together, this study suggests that the human skin fibroblasts from MERRF syndrome are subjected to higher oxidative stress resulting in more severe oxidative damage. Morever, the alteration of the expression of MMP-1 of the responses to the increased hydrogen peroxide in skin fibroblasts of MERRF patients. The alterations in gene expression imply that the disruption of mitochondrial reticulum together with degradation of structural proteins and oxidative damage to tissues may be concurrently are all involved in the pathophysiology of the MERRF syndrome.
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